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Removing unwanted protein tags recombinant bovine enterokinase REK

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Removing unwanted protein tags recombinant bovine enterokinase REK

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recombinant bovine enterokinase,For removing unwanted protein tags

Recombinant Enterokinase / Enterokinase / REK / High specificity / High purity

DESCRIPTION
Recombinant Enterokinase
YaxinBio Enterokinase is a kind of highly purified recombinant bovine enterokinase. The enzyme has been
extensively purified and there are no traces of other contaminating proteases. Enterokinase is a specific
protease that cleaves lysineC-terminal preceded by four aspartic acids: Asp-Asp-Asp-Asp-Lys. However, enterokinase does not work if this lysine is followed by a proline. Enterokinase can
remove N-terminal fusion protein,it is very useful for removing unwanted protein tags.

ADVANTAGES

High purity

(Figure 1)

(Figure 2)
Figure 1: One single main band by SDS-PAGE analysis
Figure 2: SDS-PAGE analysis of 0. 1 U EK effect on 50µg substrate after 0min, 10min, 20min, 30min, 40min
50min and 60min, respectively

High specificity

1. Protease that cleaves specifically after a lysine preceded by four aspartic acids: Asp-Asp-Asp-Asp-Lys (DDDDK)
2. No any other contaminated proteases, no non-specific cutting sites.
Conditions
Cutting condition: given an example: 25mM Tris-HCl 8.0
Fusion protein concentration: 0.1-1mg/ml (total protein content: 0.5-1.0mg)
EK content:   1-2U
Temperature:  25
Time:       overnight or 12h-16h for digestion.

Common components influence the action of enterokinase

200mM imidazole or 200mM NaCl or 5%glycerin, the reaction may be effected.The following

suggestions are given:

1) To receive the optimum result, please dialyze the sample to 25 mMTris-HCl, pH 8.0.

2) If the dialysis is inconvenient, pleasedilute the sample to 100mM imidazole, 50mMNaCl, 5%

glycerin, and the proportion of fusion protein and EK may not be changed (1U:0.5mg fusion protein).

3) If there are one or more components in samples, and cannot be removed, suggest to increase the

content of EK in reaction system or extend the reaction time.

suggestions are given:

To receive the optimum result, please dialyze the sample to 25 mMTris-HCl, pH 8.0.

If the dialysis is inconvenient, pleasedilute the sample to 100mM imidazole, 50mMNaCl, 5%

glycerin, and the proportion of fusion protein and EK may not be changed (1U:0.5mg fusion protein).

If there are one or more components in samples, and cannot be removed, suggest to increase the

content of EK in reaction system or extend the reaction time.

MAIN FEATURES

Source

E.Coli

M.W.

Theoretical MW: 25,850 Da;The apparent MW on SDS-PAGE: about 27,000 Da

Specific Activity

One unit is defined as the amount of enzyme needed to cleave 0.5mg of fusion protein in
12 to16 hours to get 95% completion at 25°C in a buffer 25mMTris-HCl, pH 8.0.Substrate:
a special fusion protein.

Storage

Store at -20°C after delivery.

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